원문정보
초록
영어
Remedies toward sustainable aquaculture rely upon research that unveils the molecular mechanisms behind host immunity and their interactions with pathogens. Antiviral defense is a major innate immune response in fish. The antiviral protein GCHV-induced gene-2 (Gig2), a member of the interferon-stimulated gene (ISG), was identified and characterized from rockfish (Sebastes schlegelii). Gig2 exists in two isoforms, namely, SsGig2-I1 and SsGig2-I2, in rockfish with lengths of 163 and 223 bp, respectively. Bioinformatic analysis indicated the availability of poly (ADP-ribose) polymerase domain in both proteins, and 51.3% identity and 71.3% similarity between both isoforms were observed. The basal expression pattern revealed the highest tissue-specific expression in rockfish gills for both isoforms. The immune challenge experiment disclosed a distinctive and strong expression of each transcript in the presence of poly I:C. Both isoforms are localized in the endoplasmic reticulum. Interferon (IFN) pathway gene analysis revealed no significant upregulation of IFN related genes. Viral hemorrhagic septicemia virus (VHSV) gene expression analysis revealed strong downregulation of viral transcripts after 48 h of infection in the presence of Gig2 isoforms. Collectively, these results indicate the protective role of Gig2 in rockfish against VHSV infection and help broaden our understanding of the innate immunity of fish.
목차
1. Introduction
2. Materials and methods
2.1. Identification of Gig2 from rockfish
2.2. Rearing fish
2.3. Tissue sampling and the immune challenge experiment
2.4. Construction of the expression plasmid
2.5. Cell culture and transfection
2.6. Downstream gene expressions
2.7. Viral hemorrhagic septicemia virus (VHSV) gene expression
2.8. RNA extraction and cDNA synthesis
2.9. Quantitative real-time PCR (qPCR)
2.10. Sub-cellular localization
2.11. Statistical analysis
3. Results and discussion
3.1. In silico analysis
3.2. Distribution of Gig2 in tissues
3.3. Immune challenge experiment
3.4. Subcellular localization of Gig2
3.5. The function of PARP domain in downstream gene expression
3.6. VHSV replication in the presence of SsGig2-I1 or SsGig2-I2
4. Conclusion
References